Kanamycin Selection Plasmids

OG148 - pSF-CMV/SP6

Starting at: $371.00

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This plasmid is compatible with all of the other DNA sequences available on this site, which are all sold in the same backbone. This flexibility allows you to create any vector you require, with a variety of functions, simply by compiling the relevant sections from our product range.

Product Code: OG148

Size (bp): 4274Bacterial Antibiotic Selection: KanamycinOrigin and Compatibility: pUC high copy, derived from pBR322 Copy Number: 500-700 copies per cellPromoter: Cytomegalovirus (CMV) immediate early promoter and the bacteriophage SP6 promoter. Purpose: A versatile expression vector for use in mammalian cells and those cells expressing the SP6 bacteriophage polymerase. This vector also contains a Kanamycin resistance cassette for growth and maintenance in E.coli. When using the SP6 promoter the vector will require linearization downstream of the region to be transcribed because this vector does not contain an SP6 terminator.

Transcription Termination: This plasmid contains three alternative transcription terminators for mammalian, bacterial and bacteriophage (T7) expression. This means that only the promoter needs to be changed to alter the expression system you are using. We sell multiple promoters that can be used in each of these systems. The presence of each terminator does not reduce expression in the alternative systems.

Inserting a Gene: Each restriction site in this vector has a purpose and allows the insertion of specific pre-designed DNA sequences. To insert a gene, we recommend using any sites between (and including) the HindIII and XbaI sites. By doing this you will be able to insert most of our other products upstream, or downstream, of your gene.

Intellectual Property Status: According to our IP-friendly policy, this plasmid is sold free of reach-through rights on any derivatives you may create.

Quality Validation: This vector has been demonstrated to express reporter genes to high levels under the CMV promoter. The start codon of these reporters was positioned within the NcoI site and the stop codon was positioned within the XbaI site. Expression was validated in the A549 lung carcinoma cell line. The SP6 promoter in this vector was demonstrated to function by in vitro transcription.

Quality Sequencing Analysis: This vector has been fully sequenced using primers F1-F10. The sequences and details of these primers are available through our website.

Genetic Modifications to standard parts:

CMV promoter – an NcoI site has been ablated in the CMV promoter. This change does not reduce expression.

KanR cassette – This KanR resistance region is not a wild type sequence, it has been modified to remove all restriction sites that conflict with the SnapFast system and to retain a high level of antibiotic resistance. It has been validated in E.coli.

Restriction site notes:

Bgl2, KpnI, SwaI and PmeI each cut the vector at two sites positioned to flank the promoter, start codon, origin, and KanR, respectively.

BsgI and BseRI recognise non-palindromic DNA sequences and cleave upstream of their recognition sites, within the stop codon that is found inside the XbaI restriction site.